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副干酪乳杆菌副干酪亚种BGSJ2-8产生的胞外多糖EPS-SJ参与对肠道上皮细胞的黏附并减少大肠杆菌与Caco-2细胞的结合。

EPS-SJ Exopolisaccharide Produced by the Strain Lactobacillus paracasei subsp. paracasei BGSJ2-8 Is Involved in Adhesion to Epithelial Intestinal Cells and Decrease on E. coli Association to Caco-2 Cells.

作者信息

Živković Milica, Miljković Marija S, Ruas-Madiedo Patricia, Markelić Milica B, Veljović Katarina, Tolinački Maja, Soković Svetlana, Korać Aleksandra, Golić Nataša

机构信息

Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade Belgrade, Serbia.

Instituto de Productos Lácteos de Asturias - Consejo Superior de Investigaciones Científicas Villaviciosa, Spain.

出版信息

Front Microbiol. 2016 Mar 9;7:286. doi: 10.3389/fmicb.2016.00286. eCollection 2016.

Abstract

The aim of this study was to determine the role of an exopolysaccharide produced by natural dairy isolate Lactobacillus paracasei subsp. paracasei BGSJ2-8, in the adhesion to intestinal epithelial cells and a decrease in Escherichia coli's association with Caco-2 cells. Annotation of the BGSJ2-8 genome showed the presence of a gene cluster, epsSJ, which encodes the biosynthesis of the strain-specific exopolysaccharide EPS-SJ, detected as two fractions (P1 and P2) by size exclusion chromatography (SEC) coupled with multi-angle laser light scattering (MALLS) detection. SEC-MALLS analysis revealed that an EPS-SJ(-) mutant (EPS7, obtained by insertion mutagenesis of the glps_2198 gene encoding primary glycosyltransferase) does not produce the P2 fraction of EPS-SJ. Transmission electron microscopy showed that EPS7 mutant has a thinner cell wall compared to the EPS-SJ(+) strain BGSJ2-83 (a plasmid free-derivative of BGSJ2-8). Interestingly, strain BGSJ2-83 showed higher adhesion to Caco-2 epithelial intestinal cell line than the EPS7 mutant. Accordingly, BGSJ2-83 effectively reduced E. coli ATCC25922's association with Caco-2 cells, while EPS7 did not show statistically significant differences. In addition, the effect of EPS-SJ on the proliferation of lymphocytes in gastrointestinal associated lymphoid tissue (GALT) was tested and the results showed that the reduction of GALT lymphocyte proliferation was higher by BGSJ2-83 than by the mutant. To the best of our knowledge this is the first report indicating that the presence of EPS (EPS-SJ) on the surface of lactobacilli can improve communication between bacteria and intestinal epithelium, implying its possible role in gut colonization.

摘要

本研究的目的是确定天然乳源分离株副干酪乳杆菌副干酪亚种BGSJ2-8产生的一种胞外多糖在其黏附于肠上皮细胞以及减少大肠杆菌与Caco-2细胞结合方面的作用。对BGSJ2-8基因组的注释显示存在一个基因簇epsSJ,其编码菌株特异性胞外多糖EPS-SJ的生物合成,通过尺寸排阻色谱(SEC)结合多角度激光光散射(MALLS)检测可将其检测为两个组分(P1和P2)。SEC-MALLS分析表明,一个EPS-SJ(-)突变体(EPS7,通过对编码初级糖基转移酶的glps_2198基因进行插入诱变获得)不产生EPS-SJ的P2组分。透射电子显微镜显示,与EPS-SJ(+)菌株BGSJ2-83(BGSJ2-8的无质粒衍生物)相比,EPS7突变体的细胞壁更薄。有趣的是,菌株BGSJ2-83对Caco-2上皮肠细胞系的黏附性高于EPS7突变体。因此,BGSJ2-83有效降低了大肠杆菌ATCC25922与Caco-2细胞的结合,而EPS7未显示出统计学上的显著差异。此外,还测试了EPS-SJ对胃肠道相关淋巴组织(GALT)中淋巴细胞增殖的影响,结果表明,BGSJ2-83对GALT淋巴细胞增殖的抑制作用高于突变体。据我们所知,这是第一份表明乳酸菌表面存在EPS(EPS-SJ)可改善细菌与肠上皮之间的交流,暗示其在肠道定植中可能发挥作用的报告。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ef5/4783416/6800871dab07/fmicb-07-00286-g001.jpg

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